Cited by 183Autophagy in Disease and TherapyUbiquitin and proteasome pathwaysCellular transport and secretion

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Autophagosome proteins LC3A, LC3B and LC3C have distinct subcellular distribution kinetics and expression in cancer cell lines.

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Koukourakis, Michael I., Kalamida, Dimitra, Giatromanolaki, Alexandra, Zois, Christos E., Sivridis, Efthimios, Pouliliou, Stamatia, Mitrakas, Achilleas, Gatter, Kevin C., Harris, Adrian L.·2015·PLoS ONE

DOI 10.1371/journal.pone.0137675PMC PMC4574774

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Abstract

LC3s (MAP1-LC3A, B and C) are structural proteins of autophagosomal membranes, widely used as biomarkers of autophagy. Whether these three LC3 proteins have a similar biological role in autophagy remains obscure. We examine in parallel the subcellular expression patterns of the three LC3 proteins in a panel of human cancer cell lines, as well as in normal MRC5 fibroblasts and HUVEC, using confocal microscopy and western blot analysis of cell fractions. In the cytoplasm, there was a minimal co-localization between LC3A, B and C staining, suggesting that the relevant autophagosomes are formed by only one out of the three LC3 proteins. LC3A showed a perinuclear and nuclear localization, while LC3B was equally distributed throughout the cytoplasm and localized in the nucleolar regions. LC3C was located in the cytoplasm and strongly in the nuclei (excluding nucleoli), where it extensively co-localized with the LC3A and the Beclin-1 autophagy initiating protein. Beclin 1 is known to contain a nuclear trafficking signal. Blocking nuclear export function by Leptomycin B resulted in nuclear accumulation of all LC3 and Beclin-1 proteins, while Ivermectin that blocks nuclear import showed reduction of accumulation, but not in all cell lines. Since endogenous LC3 proteins are used as major markers of autophagy in clinical studies and cell lines, it is essential to check the specificity of the antibodies used, as the kinetics of these molecules are not identical and may have distinct biological roles. The distinct subcellular expression patterns of LC3s provide a basis for further studies.

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LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing.
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Authors: Koukourakis, M. I., Kalamida, D., Giatromanolaki, A., Zois, C. E., Sivridis, E., Pouliliou, S., Mitrakas, A., Gatter, K. C., & Harris, A. L.
Affiliations: Democritus University of Thrace (GR)
John Radcliffe Hospital (GB)
University of Oxford (GB)
Corresponding author: Koukourakis, Michael I.
ORCID: Koukourakis, Michael I.
Kalamida, Dimitra
Giatromanolaki, Alexandra
Zois, Christos E.
Sivridis, Efthimios
Pouliliou, Stamatia
Mitrakas, Achilleas
Harris, Adrian L.
Venue: PLoS ONE
Publisher: Public Library of Science (PLoS)
Published: 2015-09-17
Volume: 10
Issue: 9
Pages: e0137675
ISSN: 1932-6203 (ISSN-L)
DOI: 10.1371/journal.pone.0137675
PMC: PMC4574774
OpenAlex: W2300762239
Cited by: 183
References: 21
Citations by year: 2026: 13 · 2025: 12 · 2024: 27 · 2023: 16 · 2022: 16 · 2021: 17 · 2020: 19 · 2019: 24 · 2018: 13 · 2017: 14 · 2016: 11 · 2015: 1
FWCI: 10.15
Type: journal article
Language: en
Open access: gold
doi.org
Licence: cc-by
creativecommons.org/licenses/by/4.0/ (vor)
Locations: PLoS ONE (publishedVersion, cc-by)
Europe PMC (PubMed Central) (publishedVersion, cc-by)
PubMed Central (publishedVersion, cc-by)
PubMed (publishedVersion)
DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders: Cancer Research UK (11359, 16466)
National Institute for Health and Care Research (NF-SI-0611-10163)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Autophagy in Disease and Therapy
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
Keywords: LC3A, LC3B, LC3C, autophagosomes, autophagy, subcellular localization, nuclear localization, Beclin-1, nuclear export, nuclear import, leptomycin B, ivermectin, antibody specificity, cancer cell lines
MeSH terms: Beclin-1, Antibodies, Autophagy, Fatty Acids, Unsaturated, Gene Expression Regulation, Humans, Ivermectin, Membrane Proteins, Microtubule-Associated Proteins, Neoplasms, Macrolides, Active Transport, Cell Nucleus, RNA Interference, RNA, Small Interfering, Cell Line, Tumor, Apoptosis Regulatory Proteins, Human Umbilical Vein Endothelial Cells

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Autophagosome proteins LC3A, LC3B and LC3C have distinct subcellular distribution kinetics and expression in cancer cell lines.

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